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Construction method of mycobacterium tuberculosis fusion protein Mtb 10.4-Hsp16.3, expression method thereof, purification method thereof and application thereof

A technology of mtb10.4-hsp16.3 and Mycobacterium tuberculosis, applied in the field of fusion proteins, can solve the problems of unstable protection and achieve the effects of prolonging the protection effect, improving the protection effect and strengthening the immune effect

Active Publication Date: 2011-08-17
LANZHOU UNIVERSITY
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Problems solved by technology

[0003] The only tuberculosis vaccine that has been successfully developed at present, Bacillus Calmette-Guerin (BCG), has been widely used around the world. However, different studies have shown that the protective effect of BCG in different populations is unstable, especially for adults with pulmonary tuberculosis. The protective efficiency is between 0-80% range

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  • Construction method of mycobacterium tuberculosis fusion protein Mtb 10.4-Hsp16.3, expression method thereof, purification method thereof and application thereof
  • Construction method of mycobacterium tuberculosis fusion protein Mtb 10.4-Hsp16.3, expression method thereof, purification method thereof and application thereof
  • Construction method of mycobacterium tuberculosis fusion protein Mtb 10.4-Hsp16.3, expression method thereof, purification method thereof and application thereof

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[0040] Preferred embodiments of the present invention are described below, and it should be understood that the preferred embodiments described here are only used to illustrate and explain the present invention, and are not intended to limit the present invention.

[0041] 1. Construction, expression and purification of Mycobacterium tuberculosis fusion protein Mtb10.4-Hsp16.3

[0042] First, the Mtb10.4 and Hsp16.3 genes were amplified by PCR and inserted into the multiple cloning site of the cloning vector in sequence to construct a recombinant vector; then the above recombinant vector was used to express the fusion protein Mtb10.4-Hsp16 in Escherichia coli. 3; Finally, purify according to the molecular weight, charge and affinity of the fusion protein Mtb10.4-Hsp16.3, and obtain the fusion protein Mtb10.4-Hsp16.3 through purification. The specific steps are as follows:

[0043] 1. Construction of fusion protein Mtb10.4-Hsp16.3(MH):

[0044] (1) According to Mtb10.4 (Rv0288...

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Abstract

The invention discloses a construction method of mycobacterium tuberculosis fusion protein Mtb 10.4-Hsp16.3, an expression method thereof, a purification method thereof and application thereof. The construction method comprises the following steps: performing the PCR (polymerase chain reaction) amplification to Mtb 10.4 gene and Hsp16.3 gene, and sequentially inserting the amplified genes into a multi-clone site of a cloning vector to construct a recombinant vector; then expressing in the colibacillus; and finally purifying to obtain the fusion protein Mtb 10.4-Hsp16.3, wherein the fusion protein can be applied in a tuberculosis subunit vaccine. The invention has the advantages that the fusion protein vaccine can induce the higher cell of the animal to react with body fluid immunoreaction; the animal challenge test shows that the vaccine plays a part in immunity enhancing on the basis of the BCG (bacille calmette guerin) immunity, so that the protective effect of the BCG can be improved and prolonged; and the vaccine has no obvious side effects.

Description

technical field [0001] The invention relates to a fusion protein, in particular to a method for constructing, expressing and purifying a mycobacterium tuberculosis fusion protein Mtb10.4-Hsp16.3, and the application of the fusion protein in tuberculosis subunit vaccines. Background technique [0002] Tuberculosis is the most widely spread, longest lasting and most serious infectious disease in the world. Since the 1980s, tuberculosis has resurged, and its morbidity and mortality rank first and second among various infectious diseases. It is estimated that 2 million people die from tuberculosis infection every year; what is more serious is that about 2 billion people are in the incubation period of Mycobacterium tuberculosis infection each year, and 5-10% of them in the incubation period turn into active tuberculosis. my country is one of the countries with a high burden of tuberculosis, and the incidence rate reaches about 1‰ of the total population of the country. Due to ...

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Application Information

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IPC IPC(8): C12N15/31C12N15/70C07K19/00C07K1/20C07K1/18C07K1/16A61K39/04A61P31/06
CPCA61K39/04C07K14/34C07K2319/40C07K2319/00A61P31/06Y02A50/30
Inventor 祝秉东牛红霞胡丽娜李青王秉翔达泽蛟辛奇万艳马维民
Owner LANZHOU UNIVERSITY
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